Method of producing a growth regulator for rumen organisms in treatment of rumen dysfunctions



George B. Garner, Columbia, Mo., assignor to Nordeu Laboratories, Inc., Lincoln, Nebr., a corporation of Delaware N Drawing. Application June 16, 1955 Serial No. 516,027

4 Claims. c1. 99-2 This invention relates to a growth regulator or con-' trolling undesirable or pathogenic organisms of certain types. For example, I have found that certain microorganisms are contaminants of the rumen in ruminant animals and Pseudomonas aeruginosa and Aerobacter aerogenes are present in rumen dysfunction cases. I have discovered that normally a regulating factor is pres-- ent in rumen fluids which inhibits the growth of these undesirable organisms or contaminants and that this factor stimulates growth of desirable bacteria which pro duces the necessary enzymes for cellulose digestion.

Therefore, it is a principal object of the present invention to "provide a growth regulator in concentrationfor use in medication and promotion of the well being and growth ofruminants and other animals. It is also an object to provide a method of preparing the growth. regulator in concentration.

Other objects of the inventionare, to incorporate the growth regulator in culturemedia containing rumen microorganisms for decreasing the lag phase or adjustment time'of the rumen organisms in such culture; incorporate-the growth regulator with-dried rumen organisms to stimulate and effect maximum activity in a shorter time when administered to a ruminant animal; to provide improved rumen function in sub-clinical diseases, such as rumen dysfunction, by adding the concentrated growth regulator to the animals' rations; to provide a growth regulator in concentrated form for suppressing or inhibiting growth of pathogenic organisms which produce rumen dysfunction and other diseased conditions in ruminant and other animals; and to provide optimal condition for cellulose digestion thereby promoting growth of animals.

In carrying out the present invention, I have discovered a factor or factors in fresh rumen fluid, its extracts, corn silage, sargo silage, feeds fermented by rumen microorganisms, material obtained from fermentation barrels inoculated with rumen microorganisms, and other similar materials, which tends to inhibit microorganisms that are considered contaminants of the rumen and which increases digestion in ruminants. To my knowledge, previous workers have not reported an antibiotic-like factor in such materials, and the factor is not identical with known antibiotics or growth regulators.

One method of obtaining the growth regulator is to prepare it from rumen material taken from freshly slaughtered animals. This material is strained or pressed to remove the liquid. The liquid material is then centrifuged. The material which has been thrown down is normally utilized for further production of rumen culture as disclosed in the Muhrer application for patent on Method of Preserving Natural Rumen Microorganisms and Product Which Results Therefrom, Serial Number 349,310, filed April 16, 1953, now Patent No. 2,738,273. The supernatant material in the centrifuging step is decanted and passed through an adsorbing column of an activated carbon at pH 3.5-4.0. The activated carbon may be of the type known under the trade Sa e name Norit-A." The Norit-A adsorbs the active factor and permits the balance of the fluid to pass through.-

The adsorbing column is then eluted with 80% alcohol.--' The alcohol dissolves the factor from the adsorbing col-' umn. The alcoholic eluate may then be concentrated by removal of the alcohol by means of vacuum distilla tion to leave the product. has the unique ability to suppress the growth of path0- The product thus obtained genic or undesirable bacteria and at the same time, it stimulates the growth of desirable bacteria or thosewhich produce enzymes for cellulose digestion. The extract is' free of microorganisms and stable pH between 3.5 and 10.

The growth regulator may be used directly for treatto autoclaving at a ment of various cases of rumen dysfunction where ru-j; men contaminants are present. It has been demonstrated to be effective against Pseudomonas aeruginosa and Aerobacter aerogenes which have been isolated from ru men dysfunction cases. The growth regulator when'added to rumen products of the type including dry rumen microorganisms and cultural nutrients such as carbohydrates, nitrogen containing chemicals, and minerals particularly disclosed in the Lew- 18 E. Harris application for patent on Combination of Rumen Culture and Culture Nutrients, Serial Number 417,966, and filed March 22, 1954, now abandoned, en

' hances the value of such products.

It has been determined that the inclusion of the growth regulating-factor in the proportion of 0.1 ml. to 11 ml.- of culture medium improves the effectiveness of the g, rumen microorganisms and the lag phase is descreased. That is, the adjustment time of the rumen microorganisms is decreased, consequently, the effective activity reaches maximum in shorter time.

The growth regulator may also be used as a premix concentrate additive to various prepared feeds for rumi nants and other animals for improving feed utilization or eificiency. r

The effectiveness of the growth regulator has been demonstrated in vitro and proved in vivo. Inhibition of Microco'ccus flavus was shown by standard plate tech- I nique.

Bactericidal or bacteriostatic action against Pseudomonas aeruginosa and Aerobacter aerogenes by turbidity measurements has also been proven. Cellulose digestion in vitro was enhanced and the lag phase of washed rumen microorganisms was shortened when 0.1 ml. of the factor was added to 11 m1. of the culture medium. The effect on cellulose digestion was checked in the artificial rumen as described by Brooks, Garner, Muhrer, and Pfander (Journal Animal Science, vol. 13, pp. 758-764, 1954), modified by reducing the amounts by one-third and using Rockefeller centrifuge tubes in place of 8-inch sugar tubes.

In one instance, one group of sheep was fed a standard ration to which the growth regulator had been added while another group of sheep was fed the same stand ard ration without the growth regulator. The control group had a marked decrease in cellulose digestion while the group receiving the growth regulator had an increase in cellulose digestion and made some gains.

The regulating factor may also be obtained from silage such as corn silage, sargo silage, and other fermented Patented Apr. 5

growth regulator. The solid material of the silage is then strained or pressed to remove the liquid fermentation material. The liquid fermentation material thus obtained is then centrifuged to throw down solids. Thesupernatant material in the centrifuging step is-decanted and passed through an adsorbing column of an activated carbon at pH 3.5-4.0. The activated carbon -may' also .be of the type known under the trade name "Nor it- A. The activated carbon adsorbs the active factor and permits the balance of the fluid to pass through. The adsorbing column is then eluted with 80% alcohol The alcohol dissolves the factor from the adsorbing column.

The alcoholic .eluate may then be concentrated by re moval of alcohol by means of vacuum distillation to leave the product.

The discovery has a wide application in practical dani- Ital production because of the stimulation of desirable rumen microorganisms and their functions such as .cel,

supernatant material from the liquid; passing the super natant material in contact with activated carbon to ;ad-.

sorb the growth regulator on the activated carbon; contacting the activated carbon with alcohol to dissolve the growth regulator, and distilling off the alcohol to leave the growth regulator product.

. 2. The method of recovering a growth regulator for inhibiting undesirable organisms and stimulating growth. of desirable bacteria in the reatment of rumen dysfunction, wherein the starting material is liquid rumen; said method including removing liquid content of the rumen; centrifuging the liquid to throw down solid materials contained in the liquid; decanting supernatant material from the liquid; passing the supernatant material in contact with activated carbon to adsorb the growth regulator on the activated carbon; contacting the activated carbon with alcohol to dissolve the growth regulator, and distilling off the alcohol to leave the growth regulator product.

3. The method of recovering a growth regulatorfor inhibiting undesirable organisms and stimulating growth of desirable bacteria in the treatment of rumen dysfunction, wherein the starting material of said method is a liquid fermentation materialfrom ruminant animalfeeds.

fermented by rumen microorganisms;,said method including centrifuging the liquid fermentation material to throw down solid materials-contained in the liquid; decanting supernatant material fromthe liquid; passing the supernatant material in contact with activated carbon to adsorb the growth regulator on the activated carbon; contacting the activated carbon with alcohol to dissolve the growth regulator, and distilling off the alcohol to leave the; growth regulator product.

4. The method of recovering a growth regulator for inhibiting undesirable organisms and stimulating growth of desirable bacteria in the treatment of rumen dysfunc- 1tion, wherein the starting material of said method is a liquid fermentation material from a silage selected from corn and sargo; said method including centrifug-ingtheliquid fermentation material to throw down .solid mate-' rials contained in the liquid; decanting supernatant material from the liquid; passing the supernatant material in contact with activated carbon to adsorb the growth regulator on the activated carbon; contacting the activated carbon with alcohol to dissolve the growth regulator, and distilling off the alcohol to leave the growth.

' regulator product.

References Cited in the file of this patent I UNITED STATES PATENTS 2,530,416 Wolf Nov. 21, 1950 2,563,794 Rickes Aug. 7, 1951 2 ,646,386 Miner July 21, 1953 2,700,611 Jeifreys Jan. 25, 19,55

OTHER REFERENCES V Burroughs et all: Jour. of Animal Science, 513, 1950. 

1. THE METHOD OF RECOVERING A GROWTH REGULATOR FOR INHIBITING UNDESIRABLE ORGANISMS AND STIMULATING GROWTH OF DESIRABLE BACTERIA IN THE TREATMENT OF RUMEN DYSFUNCTION, WHEREIN THE STARTING MATERIAL OF SAID METHOD IS A FERMENTATION PRODUCT OF THE CLASS INCLUDING LIQUID RUMEN, LIQUIDS OF CORN AND SARGO SILAGES, AND ANIMAL FEEDS FERMENTED BY RUMEN MICROORGANISMS, SAID METHOD INCLUDING CENTRIFUGING THE LIQUID FERMENTATION MATERIAL TO THROW DOWN SOLID MATERIALS CONTAINED IN THE LIQUID, DECANTING SUPERNATANT MATERIAL FROM THE LIQUID, PASSING THE SUPERNATANT MATERIAL IN CONTACT WITH ACTIVATED CARBON TO ADSORB THE GROWTH REGULATOR ON THE ACTIVATED CARBON, CONTACTING THE ACTIVATED CARBON WITH ALCOHOL TO DISSOLVE THE GROWTH REGULATOR, AND DISTILLING OFF THE ALCOHOL TO LEAVE THE GROWTH REGULATOR PRODUCT. 